Comparison And Analysis Of The Constituents Of Lithospermum Erythorhizon And Soft Root From Different Origin

Objective1.To optimize the ultrasound extraction technology of naphthalene quinone in Zicao.2.To establish a method for determination of L-shikonin,acetylshikonin,?-aceto-xyisovaleryalkannin,deoxyshikonin,isobutylshikonin and?,??-dimethylacrylalkannin in Zicao.Compare the difference of six quinones in Lithospermum erythorhizon and the soft roots from different habitats.3.To establish an HPLC fingerprint for Lithospermum erythorhizon.To analyze the constituents of Lithospermum erythorhizon and the soft roots from different habitats.Method1.An L9?3⁴?orthogonal experiment was adopted to investigate the three factors?solvent of extraction,time of extraction and solid-liquid ratio?,with the content of naphthalene quinone as indexes for evaluation.Each factor was selected at 3 levels for testing.2.HPLC method was used as follow:The separation was carried out on Symmetry C18?4.6 mm×250 mm,5?m?chromatographic column with mobile phase consisted of acetonitrile-water?70:30,V/V?at the flow rate of 1.0 mL·min^-1.The detection wavelength was set at 516 nm and thecolumn temperature was 30?.3.The extract of Lithospermum erythorhizon?Arnebia euchroma and Anhui puccoon were prepared.HPLC fingerprints were established.Results1.Solvent of extraction,time of extraction and solid-liquid ratio were the three factors.The solvent of extraction had the greatest effect on the extraction of naphthalene quinone in Zicao.The content of each component was the highest when acetonitrile:water?70:30?as the solvent of extraction.2.The linear ranges of L-shikonin,acetylshikonin,?-acetoxyisovaleryalkannin,deoxy-shikonin,isobutylshikonin and?,??-dimethylacrylalkannin were 2.40²4?g·mL^-1?r=0.9992?,9.6⁹6?g·mL^-1?r=0.9992?,38³80?g·mL^-1?r=0.9998?,2.2²2?g·mL^-1?r=0.9993?,8.5⁸5?g·mL^-1?r=0.9996?and 3.4³4?g·mL^-1?r=0.9994?,respectively.The average recoveries were 96.02%?RSD=2.55%,n=6?,103.38%?RSD=2.72%,n=60?,98.34%?RSD=1.74%,n=6?,99.00%?RSD=2.29%,n=6?,88.83%?RSD=2.40%,n=6?and 87.82%?RSD=1.67%,n=6?.The contents of L-shikonin,acetylshikonin,deoxyshikonin,isobutylshikonin and?,??-dimethy-lacrylalkannin in Lithospermum erythorhizon were 0.010%,0.103%,0.0097%,0.090%,0.0122%;the contents of L-shikonin,acetylshikonin,?-acetoxyisovaleryalkannin,deoxyshik-onin,isobutylshikonin and?,??-dimethylacrylalkannin in Arnebiaeuchroma were 0.025%,0.152%,0.886%,0.0110%,0.123%,0.0667%;the contents of L-shikonin,acetylshikonin,?-acetoxyisovaleryalkannin,deoxyshikonin,isobutylshikonin and?,??-dimethylacrylalkan-nin in Anhui puccoon were 0.029%,0.178%,0.616%,0.0103%,0.167%,0.0632%.3.Under the above chromatographic conditions,the mutual mode of HPLC fingerprints was set up.Seven common peaks were selected as the common fingerprint peaks in Lithospermum erythorhizon,eight common peaks were selected as the common fingerprint peaks in Arnebia euchroma and Anhui puccoon.HPLC fingerprints of Zicao showed good precision,stability and reproducibility;the similarity of the fingerprints in nine batches of Zicao was more than 0.950.Conclusions1.Taking all factors into consideration,the extraction process was determined:using acetonitrile-water?70:30,V/V?,time of ultrasonic extraction was 40min and solid-liquid ratio was 1:40.The ultrasonic extraction process was stable and convenient.2.Develop a method for simultaneous determination of six quinones in Lithospermum erythorhizon and the soft roots from different habitats.In Lithospermum erythorhizon,five kinds of quinones were detected,including L-shikonin,acetylshikonin,deoxyshikonin,isobutylshikonin and?,??-dimethylacrylalkannin.?-acetoxyisovaleryalkannin was not detected.There were differences in the content of the quinone components between Lithospermum erythorhizon and the soft roots from different habitats.The content of six quinones in Arnebia euchroma and Anhui puccon was higher,which shown that genuine medicinal herbs had their scientific characteristics.Lithospermum erythorhizon had same medicinal ingredients with genuine medicinal herbs and had certain medicinal value.3.There were eight common peaks in the HPLC fingerprints of Arnebia euchroma and Anhui puccon,there were seven common peaks in Lithospermum erythorhizon,?-acetoxyisovaleryalkannin was not detected.HPLC fingerprints of Lithospermum erythorhizon,Arnebia euchroma and Anhui puccon had the same chromatographic peaks,which further confirmed that Lithospermum erythorhizon had certain medicinal value.